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rhBMP-2 壳聚糖纳米缓释载体-负载 PLGA/nHA 支架构建下颌组织工程骨的实验研究。

Experimental study of rhBMP-2 chitosan nano-sustained release carrier-loaded PLGA/nHA scaffolds to construct mandibular tissue-engineered bone.

机构信息

Department of Oral & Maxillofacial Surgery, Affiliated Hospital of Qingdao University, Qingdao, Shandong, China; School of Stomatology, Qingdao University, Qingdao, Shandong, China.

Department of Oral & Maxillofacial Surgery, Affiliated Hospital of Qingdao University, Qingdao, Shandong, China; School of Stomatology, Qingdao University, Qingdao, Shandong, China; Dental Digital Medicine & 3D Printing Engineering Laboratory of Qingdao, Qingdao, Shandong, China; Shandong Provincial Key Laboratory of Digital Medicine and Computer-assisted Surgery, Qingdao, Shandong, China.

出版信息

Arch Oral Biol. 2019 Jun;102:16-25. doi: 10.1016/j.archoralbio.2019.03.023. Epub 2019 Mar 28.

Abstract

OBJECTIVES

To develop PLGA/nHA scaffold containing BMP-2 cell growth factor chitosan sustained release system as tissue engineered bone for repairing large jaw defects, test its sustained release rhBMP-2 efficiency in vitro, and evaluate its Osteogenesis in rabbit mandibular defects.

METHODS

Tissue engineered bone scaffold complexes were prepared by complexing rhBMP-2 loaded chitosan (CS / rhBMP-2) nano sustained release carrier with PLGA / nHA scaffold carrier by 3D printing. In vitro, the porosity, pore size and degradation rate and the dose-time effect relationship of cytokine release were examined. Micro-CT, hematoxylin/eosin staining, Masson staining and immunohistochemistry were determined at week 4, week 8 and week 12 in 18 rabbits.

RESULTS

In vitro, the porosity was (73.64 ± 1.82)%, and the average pore diameter was (431.31 ± 18.40) μm. The cumulative release was only 9.54 ± 0.86% within 48 h and 61.38 ± 2.39% on the 30th day. In vivo, Micro-CT examination showed that the BMD and the bone volume fraction at 4, 8, and 12 weeks were higher in the implantation group than in the control group. In the 12th week, Masson staining showed that new bone occupied 19.2% of the defect area in the control group, whereas the proportion of new bone reached 45.5% in the experimental group.

CONCLUSIONS

PLGA/nHA/CS/rhBMP-2 scaffold complex effectively controlled the early burst effect of rhBMP-2. The bone tissue engineering scaffold complex had good biocompatibility and induced osteogenic effects. It successfully repaired the experimental bone defect area of the rabbit mandible.

摘要

目的

构建载 BMP-2 细胞生长因子壳聚糖的 PLGA/nHA 支架作为组织工程骨用于修复大的颌骨缺损,检测其在体外对 rhBMP-2 的持续释放效率,并评价其在兔下颌骨缺损中的成骨作用。

方法

通过 3D 打印技术将负载 rhBMP-2 的壳聚糖(CS/rhBMP-2)纳米缓释载体与 PLGA/nHA 支架载体复合,制备组织工程骨支架复合物。体外考察支架复合物的孔隙率、孔径、降解率以及细胞因子释放的量效关系。18 只兔分别于 4、8、12 周时行 Micro-CT 检测、苏木精-伊红(HE)染色、Masson 染色及免疫组织化学染色。

结果

体外实验中,支架复合物的孔隙率为(73.64±1.82)%,平均孔径为(431.31±18.40)μm。48 h 内累计释放量仅为 9.54±0.86%,第 30 天为 61.38±2.39%。体内实验中,Micro-CT 检查显示,植入组在第 4、8、12 周时的骨密度和骨体积分数均高于对照组。第 12 周时,Masson 染色显示对照组缺损区新生骨占 19.2%,实验组则达到 45.5%。

结论

PLGA/nHA/CS/rhBMP-2 支架复合物有效控制了 rhBMP-2 的早期突释效应。该骨组织工程支架复合物具有良好的生物相容性,能诱导成骨作用,成功修复了兔下颌骨的实验性骨缺损区。

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